College of Horticulture, State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, China.
State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei, China.
Plant Biotechnol J. 2019 Sep;17(9):1770-1787. doi: 10.1111/pbi.13099. Epub 2019 Mar 19.
WRKY comprises a large family of transcription factors in plants, but most WRKY members are still poorly understood. In this study, we report the identification and functional characterization of PbrWRKY53 isolated from Pyrus betulaefolia. PbrWRKY53 was greatly up-regulated by drought and abscisic acid, but slightly induced by salt and cold. Subcellar localization analyses showed that PbrWRKY53 was located in the nucleus. Ectopic expression of PbrWRKY53 in tobacco and Pyrus ussuriensis conferred enhanced tolerance to drought stress. The transgenic plants exhibited better water status, less reactive oxygen species generation and higher levels of antioxidant enzyme activities and metabolites than the wild type. In addition, overexpression of PbrWRKY53 in transgenic tobacco resulted in enhanced expression level of PbrNCED1, and led to the increase in larger amount of vitamin C accumulation in comparison to WT. Knock-down of PbrWRKY53 in P. ussuriensis down-regulated PbrNCED1 abundance, accompanied by compromised drought tolerance. Yeast one-hybrid assay, EMSA and transient expression analysis demonstrated that PbrWRKY53 could bind to the W-box element in the promoter region of PbrNCED1. Taken together, these results demonstrated that PbrWRKY53 plays a positive role in drought tolerance, which might be, at least in part, promoting production of vitamin C via regulating PbrNCED1 expression.
WRKY 包含植物中一个大型转录因子家族,但大多数 WRKY 成员的功能仍知之甚少。本研究从梨中分离到 PbrWRKY53,并对其进行了鉴定和功能分析。PbrWRKY53 受干旱和脱落酸强烈诱导,但受盐和冷胁迫的轻微诱导。亚细胞定位分析表明 PbrWRKY53 定位于细胞核。在烟草和砂梨中的异位表达增强了其对干旱胁迫的耐受性。与野生型相比,转基因植物表现出更好的水分状态、更少的活性氧生成、更高的抗氧化酶活性和代谢物水平。此外,在转基因烟草中过表达 PbrWRKY53 导致 PbrNCED1 的表达水平升高,与 WT 相比,维生素 C 的积累量也增加。在砂梨中敲低 PbrWRKY53 会降低 PbrNCED1 的丰度,同时导致其耐旱性下降。酵母单杂交实验、EMSA 和瞬时表达分析表明,PbrWRKY53 可以结合 PbrNCED1 启动子区域的 W-box 元件。总之,这些结果表明 PbrWRKY53 在耐旱性中发挥积极作用,至少部分是通过调节 PbrNCED1 表达促进维生素 C 的产生。